Fibroblast cells stained for mitochondria and nuclei. 20x air (0.7 NA) Leica widefield image of living human fibroblast cells stained for mitochondria (Mitotracker green, Invitrogen Inc.) and nuclei (Hoechst). Z stack fluorescent channels were deconvolved in Huygens Essential, then the mitochondrial channel was processed in Object Analyzer to identify the number of mitochondrial objects per cell. Resulting objects were separately color coded and overlaid with a maximum projection of the nuclei, and the in focus DIC image.

Image created by Dr. Glyn Nelson, University of Newcastle upon Tyne, UK

The Power of Deconvolution. Experimental point spread functions were generated for the red, green, and blue channels on an epifluorescence microscope and then used to deconvolve a standard Invitrogen Floucells #1 prepared slide, containing bovine pulmonary artery endothelial cells stained for mitochondria (red), F-actin (green), and nuclei (blue). Before (left) and after (right) deconvolution images were merged side by side to display the power of deconvolution.

Inhibitory axon terminals at the pyramidal layer of CA1 area of the hippocampus. A subset of terminals is immunostained for parvalbumin (blue), while another population is labeled with CB1 cannabinoid receptor (green) and VGluT3 vesicular glutamate transporter (red). The receptor is on the cell membrane while vesicular transporter is present intracellularly. Slice view of a Huygens deconvolved confocal stack from a Nikon Ti-E, using Nikon A1R 4 channel detector unit.

Detail of an imaginal disc from a third instar Drosophila Melanogaster larva.Left: a slice of the original data, imaged using an Andor Revolution spinning disc confocal microscope.Right: the same slice, deconvolved using Huygens Professional. The fixed sample was stained against alfa-tubulin (green) and gamma-tubulin (red).

Recorded by Dr. Paula Sampaio, Advanced Light Microscopy Facility, University of Porto.

Gamma tubulin fiber structure in an imaginal disc.Left: SFP rendering of the original data, imaged using an Andor Revolution spinning disc confocal microscope. Right: SFP rendering of the same data, deconvolved using Huygens Professional.

Recorded by Dr. Paula Sampaio, Advanced Light Microscopy Facility, University of Porto.

Jurkat T-cell making contact with a Raji B-cell, stained for Raji cytosol (FURA-2, blue), T-cell receptor (anti-CD3 Alexa647, green), and Actin (Lifeact-mRFPruby, red)Left: Maximum intensity projections (XZ, XY, ZY) of the original data, imaged using a Zeiss Cell Observer HS system at 37ºC with a 40x Fluor 1.3 oil lens. This is a single frame from a time lapse acquisition. Right: Deconvolved using Huygens Essential. In the time series a ring-like pattern of Actin and a bulls-eye pattern of CD3 can be seen, accumulating at the contact-site (the immunolgical synapse). Both structures gave us a hint that the T-cell was activated properly."

Recorded by Christian Junker M.Sc., Institut für Biophysik, University of Saarland, Germany.

Macrophage fluorescently stained for tubulin (yellow), actin (red) and the nucleus (DAPI, blue).Left: original image, recorded with a wide field microscope. Right: the same dataset, deconvolved using Huygens Professional. The datasets are visualized with top-view maximum intensity projections.

Cell nuclei labelled with Draq5 (XY slice)Left: a slice of the original data, imaged using an inverted Leica TCS SP2 AOBS confocal microscope. Right: the same slice, deconvolved using Huygens Professional. This image was used to test an automatic nuclei counting algorithm. Huygens deconvolution enhances the performance of this method. %% Recorded by Dr. Nicolas Fête, Laboratory of Stem Cells Dynamics, Swiss Federal Institute of Technology of Lausanne.

Cell nuclei labelled with Draq5 (axial XZ slice)Left: a slice of the original data, imaged using an inverted Leica TCS SP2 AOBS confocal microscope. Right: the same slice, deconvolved using Huygens Professional. This image was used to test an automatic nuclei counting algorithm. Huygens deconvolution enhances the performance of this method.

SFP rendering of a cell cluster. The XY and XZ slices in the previous images are part of this dataset. In the SFP volume rendering algorithm the data is taken as a distribution of fluorescent dye. By modeling a physical light/matter interaction process an image is computed showing the data as it would have appeared in reality when viewed under these conditions.

The original data is recorded by Dr. Nicolas Fête, Laboratory of Stem Cells Dynamics, Swiss Federal Institute of Technology of Lausanne.

Axial image of an isolated rat Hepatocyte couplet.Left: an XZ slice of the original confocal 3D image. Right: the same XZ slice after restoring the whole stack with the Huygens Software. The data shows two adhering liver cells stained with phalloidin for actin (red), tubulin (blue) and dextran as a marker for endocytosis (green).

Data recorded by Dr. Permsin Marbet at the Department of Anatomy, University of Basel, Switzerland, in the lab of Prof. Lukas Landmann. (See MarbetHepatocyte for more details).

4Pi two-photon image of F-actin filaments of a mouse skin fibroblast cell. Left: a 'conventional' confocal image. Right: the restored 4Pi two-photon image. Both 3D-images were recorded at the same site of the specimen to allow a direct comparison of both methods. The F-actin fibers are directed along the Y-axis, i.e., perpendicular to the axial image. The 4Pi-confocal restored image reveals more details of the object than the confocal counterpart as a result of the improved 3D-resolution. The actin fiber in the center is substantially better resolved. The axial FWHM resolution in the restored 4Pi image was shown to be 70nm.

Image created by Dr. Barbara Rothen, Institute of Anatomy, University of Bern, Switzerland

BEM-1-GFP at septal pore of N. crassa hypha. The function of BEM-1 in Neurospora crassa is still unkown. The bottom right picture shows a 3D-reconstruction of the septal pore which is rotated about 90°. Bright/Wide-field image stacks were recorded with Zeiss Examiner.D1 with 100X/1.30NA oil objective, and deconvolved with Huygens.

Images were recorded by Dr. Timo Schuerg at the Department of Genetics, TU Braunschweig, Germany

Trypanosoma cruzi-infected Vero cells. Vero cells infected with T. cruzi were labeled with two monoclonal antibodies: 3B2 (shown in green) reacts with the flagellated trypomastigote forms, and 4B5 (in red) recognizes an intracellular structure in all parasite forms. DAPI (in blue) was used to stain all DNA rich organelles - kinetoplasts and nuclei. Serial Z sections acquired on a BioRad confocal microscope were deconvolved and SFP-rendered using Huygens.

Multicolored bead image. Compilation of three 500nm beads (in 2 colors) and one 200nm bead in four colors. Image was rendered with the Huygens Simulated Fluorescence Process Renderer. Note that the chromatic shift is clearly visible.

Mitochondrial dysfunction in senescent fibroblasts. 20x air (0.7 NA) widefield image captured on a Leica upright of living human fibroblast cells stained to show the mitochondrial network (green) and it's activity (red) and cell nuclei (blue, using mitotracker green, TMRM and Hoechst respectively). Z stack fluorescent channels were deconvolved in Huygens Essential and rendered using SFP. Individual cells can be seen to have varying levels of energy production (red intensity), as well as intracellular variation between individual mitochondrial networks.

Image created by Dr. Glyn Nelson, University of Newcastle upon Tyne, United Kingdom

Chromatic shift between blue and red channel. A chromatic shift between blue and red stained centromeric satellite repeat regions can be clearly distinguished. Chromatic shift can be corrected by using the Huygens Chromatic Shift Corrector.